The Use of Fluorescent Probes to Detect ROS in Photodynamic Therapy

Photodynamic therapy employs nontoxic dyes called photosensitizers (PS) that are excited by visible light of the correct wavelength to produce a variety of reactive oxygen species (ROS) by an interaction between the long-lived PS triplet states with ambient oxygen. The most important type of ROS in photodynamic therapy (PDT) is singlet oxygen, which is produced by a Type II energy transfer process. On the other hand, superoxide, hydrogen peroxide, and hydroxyl radicals can be produced by a Type I electron transfer process. This chapter describes a set of fluorescent probes that can be used to tease apart these different ROS produced when various PS are illuminated in solution. Singlet oxygen sensor green (SOSG) is used for singlet oxygen, 4-hydroxyphenyl-fluorescein (HPF) for hydroxyl radicals, Amplex Red for hydrogen peroxide, and nitroblue-tetrazolium or XTT for superoxide.